Process for preparation of improved mutant strain of...

C - Chemistry – Metallurgy – 12 – N

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167/135, 195/1.3

C12N 1/20 (2006.01) A61K 39/10 (2006.01)

Patent

CA 1315226

ABSTRACT OF THE DISCLOSURE The present invention provides a process for preparation of a mutant strain of B. bronchiseptica having chromosomal genetic markers carrying at least temperature-sensitive (grown at 42°C), nalidixic acid- resistant and heat-labile dermonecrotic toxin producing ability negative markers preferable for preparation of live attenuated AR vaccine, which comprises subjecting a naturally isolated parent, wild or virulent strain of B. bronchiseptica, to growth at 42°C in the presence of nalidixic acid on a Bordet-gengou agar plate to induce mutation, isolating a mutant strain having the genetic markers of being at least temperature-sensitive (grown at 42°C), nalidixic acid-resistant and heat-labile dermonecrotic toxin negative, culturing the mutant strain of B. bronchiseptica resulting from the induced mutation on a Bordet-gengou agar plate containing nalidixic acid at 42°C, thereafer picking up an organism morphologically resembling a phase-I organism, and repeating the procedures of cultivation on the Bordet-gengou agar plate and picking up phase-I organisms. Also provided by the invention is a live attenuated AR vaccine for prophylaxis of B. bronchiseptica infection, which comprises B. bronchiseptica phase-I organisms of strain B-42 having chromosomal genetic markers of being temperature-sensitive (grown at 42°C), nalidixic acid-resistant and heat- labile dermonecrotic toxin negative.

550904

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